Elliptic Fourier Analysis of leaf outline shape in forest fragment populations of <Emphasis Type="Italic">Anthurium sinuatum</Emphasis> and <Emphasis Type="Italic">A. pentaphyllum</Emphasis> (Araceae) from Northeast Brazil

A field-based comparative study of leaflet outline shape in five populations of Anthurium sinuatum Benth. ex Schott from Amazonia and humid brejo forests of Ceará, Northeast Brazil, and ten populations of A. pentaphyllum (Aubl.) G. Don var. pentaphyllum from Amazonia and the Brazilian Atlantic forest is reported. 1,120 leaflets from 561 individuals in 15 populations were sampled. Using images, leaflet outlines were captured by digitisation and subjected to Elliptic Fourier Analysis (EFA) to produce matrices of Fourier coefficients based on 40 harmonics; lateral and central leaflets were analysed separately. Twelve shape variables, extracted from the Fourier coefficient matrices, described the leaflet outlines of 531 individuals. Between-population relationships were estimated using cluster analysis, Canonical Variates Analysis and non-parametric MANOVA. Amazonian populations showed the greatest within-population variability. In A. sinuatum the Pacatuba population was the most distinct in Ceará and the Pará population (Amazonia) was not significantly different from those at Ibiapaba and Maranguape (both Ceará). In A. pentaphyllum 80% of population pairs were significantly different and three significantly different groups of populations were observed. Although the populations of A. pentaphyllum displayed considerable variation, the morphological patterns did not correlate with previously reported molecular ones. The two species were found to be slightly but significantly different on leaf outline data.     

Morphological variation in leaf shape in <Emphasis Type="Italic">Ainsliaea apiculata</Emphasis> with special reference to the endemic characters of populations on Yakushima Island,Japan

We analyzed leaf shape variations in Ainsliaea apiculata Sch. Bip. to evaluate the uniqueness of morphological characters in populations on Yakushima Island, Kagoshima Prefecture, Japan. Leaf size and shape from populations on Yakushima Island (n = 300) were compared with those from populations in other areas of Japan (n = 300). A considerable amount of variation occurred in leaf size in A. apiculata populations both on Yakushima Island and elsewhere, but clear discontinuities in leaf size were not detected. Some variants previously thought to be endemic to Yakushima Island, i.e., A. apiculata var. acerifolia and A. apiculata var. rotundifolia, were also found in other locations in Japan. Moreover, these leaf types were found to be continuous with the typical leaf shape of A. apiculata var. apiculata via various intermediate types, suggesting the need for future revision of these taxa. Based on these results, we reevaluated the uniqueness of the Yakushima populations of A. apiculata in terms of leaf variation. The uniqueness of the Yakushima populations was defined by a more diverse leaf shape than found in populations from other areas.     

A preliminary study of genetic variation in populations of Monstera adansonii var. klotzschiana (Araceae) from North-East Brazil, estimated with AFLP molecular markers

BACKGROUND AND AIMS: This study sought genetic evidence of long-term isolation in populations of Monstera adansonii var. klotzschiana (Araceae), a herbaceous, probably outbreeding, humid forest hemi-epiphyte, in the brejo forests of Ceará (north-east Brazil), and clarification of their relationships with populations in Amazonia and the Atlantic forest of Brazil. METHODS: Within-population genetic diversity and between-population dissimilarity were estimated using AFLP molecular markers in 75 individuals from eight populations located in Ceará, the Brazilian Atlantic Forest and Amazonia. KEY RESULTS: The populations showed a clinal pattern of weak genetic differentiation over a large geographical region (F(ST) = 0.1896). A strong correlation between genetic and geographical distance (Mantel test: r = 0.6903, P = 0.002) suggests a historical pattern of isolation by distance. Genetic structure analysis revealed at least two distinct gene pools in the data. The two isolated Ceará populations are significantly different from each other (pairwise Phi(PT) = 0.137, P = 0.003) and as diverse (Nei's gene diversity, average H(e) = 0.1832, 0.1706) as those in the Atlantic and Amazon forest regions. The population in southern Brazil is less diverse (Nei's gene diversity, average H(e) = 0.127) than the rest. The Ceará populations are related to those of the Atlantic forest rather than those from Amazonia (AMOVA, among-groups variation = 11.95 %, P = 0.037). CONCLUSIONS: The gene pools detected within an overall pattern of clinal variation suggest distinct episodes of gene flow, possibly correlated with past humid forest expansions. The Ceará populations show no evidence of erosion of genetic diversity, although this was expected because of their isolation. Their genetic differentiation and relatively high diversity reinforce the importance of conserving the endangered brejo forests.     

Genetic variation in natural populations of Anthurium sinuatum and A. pentaphyllum var. pentaphyllum (Araceae) from north‐east Brazil using AFLP molecular markers

Genetic variation was investigated using AFLP markers in 12 populations of Anthurium sinuatum and A. pentaphyllum var. pentaphyllum (Araceae) in north‐east Brazil, Amazonia and the Brazilian Atlantic forest. Two unique genetic patterns characterized the populations of A. sinuatum as a group, but no correlation between genetic and geographical interpopulation distance was found; the Amazonian population was not separated from that in Ceará. The isolated Ceará brejo populations of A. sinuatum were genetically distinct, but genetic diversity levels were similar to populations elsewhere, with no evidence of genetic erosion. Anthurium pentaphyllum populations were significantly different from each other; Bayesian genetic structural analysis found no common genetic pattern, but revealed genetic clusters unique to subgroups and individual populations in the Atlantic forest and French Guiana. Anthurium pentaphyllum and A. sinuatum can be distinguished genetically, but individuals of both species formed intermediate genetic clusters that blurred their distinction. We suggest that genetic mixing of A. sinuatum and A. pentaphyllum has occurred in north‐east Brazil, possibly connected with cycles of humid forest expansion. The weak genetic structure in A. sinuatum is consistent with the natural fragmentation of continuous forest areas, possibly during the Holocene. This study highlights the scientific importance of the highly threatened brejo forests for tropical American biogeography. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 159 , 88–105.     

Morphological and molecular variation in Mitchella undulata,with special reference to the systematic treatment of the dwarf form from Yakushima

Morphological and molecular variation in Mitchella undulata Siebold et Zucc. was examined to evaluate the genetic basis for recognizing the dwarf variety, M. undulata var. minor Masamune. Considerable variation in leaf size in M. undulata, but no obvious morphological discontinuities, were found between the normal and dwarf varieties. Instead, a weak cline running from the Pacific Ocean to the Sea of Japan was found. Anatomical observations of leaf blades revealed that the large variation in leaf size can be attributed to variation in the number of leaf cells and not to differences in cell size. A molecular analysis based on sequences of rDNA internal transcribed spacer regions indicated that there were two major genotypes in M. undulata with minor variation in haplotypes resulting from additional substitutions or putative recombination. The dwarf form from Yakushima was neither genetically uniform nor apparently differentiated from other populations. From these results, we conclude that the dwarf form of M. undulata should be treated at the rank of forma.     

Spatial genetic structure within two populations of a self-incompatible perennial,Chionographis japonica var.japonica (Liliaceae)

Intrapopulational spatial genetic structure was examined in two populations ofChionographis japonica var.japonica, a self-incompatible perennial, by spatial autocorrelation analysis of enzyme polymorphism. Although most spatial autocorrelation indices (Moran'sI) in the shortes distance class were significantly positive, most in the other distance classes did not significantly deviate from the values expected from random distributions of genotypes in both populations. This contrasts with a spatial genetic pattern previously reported for a population of the predominantly selfing congener,C. japonica var.kurohimensis, indicating that pollen-mediated gene flow highly impedes genetic substructuring within populations of outcrossingC. japonica var.japonica. Genetic similarity in very proximate distance found in outcrossingC. japonica var.japonica is probably due to restricted dispersal of seeds.     

Early effects of the mutation laurina on the functioning and size of the shoot apex in coffee tree and analysis of the plastochron phases: relationships with the dwarfism of leaves

The current article presents the investigations into the effect of the laurina mutation on the functioning and size of the shoot apical meristem (SAM) in Coffea arabica. This monolocus and Mendelian mutation is known to have pleiotropic effects on tree shape and dwarfism. A comparison between the wild type C. arabica var. Bourbon and its natural dwarf mutant C. arabica var. laurina, also called Bourbon pointu, was carried out leading to three main results: (1) the effects appeared immediately after the emergence of the buttress but did not affect the dome-shaped SAM (size and shape); (2) the effects were located at the peripheral zone and maintained subsequently within the leaf primordia; (3) the effects consisted of reduction in both the size of primordia and the height of incipient internode, consequently resulting in dwarfism of mature leaves and internodes. By contrast, the laurina mutation had no effect on the relationship between the phyllochron and the plastochron, the decussate and opposite phyllotaxis, and the relative timing of SAM functioning within the plastochron.     

Taxonomic status of Monotropastrum humile, with special reference to M. humile var. glaberrimum (Ericaceae, Monotropoideae)

Taxonomic treatment of the achlorophyllous monotropoid plant Monotropastrum humile is still unclear and confusing because of the lack of detailed morphological analyses and molecular phylogeny. In particular, the taxonomic status of a glabrous variety, M. humile var. glaberrimum, is under debate. Our detailed examination of the morphological characteristics of living plants revealed that M. humile var. glaberrimum can be easily distinguished from the putative conspecific taxon M. humile var. humile by characteristics not previously recognized, namely the shape and color of the floral disc. Most morphological features characterizing Cheilotheca were also found in M. humile var. glaberrimum. Moreover, there was considerable nucleotide differentiation in the internal transcribed spacer (ITS)2 sequences of M. humile var. humile and var. glaberrimum. Molecular analysis of the phylogenetic relationship of M. humile var. humile, var. glaberrimum, and other monotropoids using ITS2 sequences showed that two varieties of M. humile formed a monophyletic clade with a member of a different genus, Monotropa L., but obvious phylogenetic relationships among these three taxa were not obtained. Thus we conclude that Monotropastrum humile var. glaberrimum should be treated as a distinct species. However, the generic affiliation of M. humile var. glaberrimum could not be determined because of its intermediate character state combination and the insufficient characterization of related species. We strongly suggest that Monotropastrum as a whole needs re-evaluation.     

A taxonomic study of KoreanSmilacina

Several taxonomic problems of KoreanSmilacina were examined by statistical methods, based on morphological data. Consequently,S. davurica described in Korean flora differs greatly from originalS. davurica, and closely related toS. bicolor morphologically. Therefore, the present authors treated this new taxon as a variety ofS. bicolor, S. bicolor var.flavovirens N. S. Lee et J. Y. Kim var. nov (“Yondusomdae” in Korean). The distribution in South Korea ofS. bicolor var.bicolor is known as only Mt. Chiri, but this species is collected in Mt. Dukyou, also. The bounds of morphological variations ofS. japonica var.japonica, the short plant size and no hairs on the stem, were revealed. The morphological characteristics ofS. japonica var.japonica andS. japonica var.mandshurica is obscure to distinguish. According to the present study,S. japonica var.mandshurica differs fromS. japonica var.japonica in plant size, size of adaxial leaf epidermal cell, ratio of style/ovary length and shape of stigma. The unrecorded taxon in Korean flora,S. robusta, was given a new Korean name as “Kunsomdae.” AlthoughS. japonica var.mandshurica is the variety ofS. japonica var.japonica, it is more closely related toS. robusta than toS. japonica var.japonica.     

Construction of genetic linkage map of the medicinal and ornamental plant <Emphasis Type="Italic">Catharanthus roseus</Emphasis>

An integrated genetic linkage map of the medicinal and ornamental plant Catharanthus roseus, based on different types of molecular and morphological markers was constructed, using a F₂ population of 144 plants. The map defines 14 linkage groups (LGs) and consists of 131 marker loci, including 125 molecular DNA markers (76 RAPD, 3 RAPD combinations; 7 ISSR; 2 EST-SSR from Medicago truncatula and 37 other PCR based DNA markers), selected from a total of 472 primers or primer pairs, and six morphological markers (stem pigmentation, leaf lamina pigmentation and shape, leaf petiole and pod size, and petal colour). The total map length is 1131.9 cM (centiMorgans), giving an average map length and distance between two markers equal to 80.9 cM and 8.6 cM, respectively. The morphological markers/genes were found linked with nearest molecular or morphological markers at distances varying from 0.7 to 11.4 cM. Linkage was observed between the morphological markers concerned with lamina shape and petiole size of leaf on LG1 and leaf, stem and petiole pigmentation and pod size on LG8. This is the first genetic linkage map of C. roseus.     

自然干旱梯度下的酸枣表型变异

表型变异是植物应对环境变化的一种策略。酸枣植物从中国东部沿海到内陆腹地均有分布,其表型性状的变异可能解释其对自然干旱梯度的适应机制。为验证这一假说,以烟台、石家庄、银川、吐鲁番4个自然干旱梯度生境中生长的酸枣三年生植株的12个表型性状为调查研究对象,采用变异系数和巢式方差分析对酸枣的表型变异进行分析。结果表明:(1)从烟台到银川,叶面积、叶长、叶周长和叶柄长总体呈减小的趋势,而比叶面积呈增大的趋势;(2)随着干旱程度的增强,二次枝的长度、二次枝的基部粗、二次枝的枣吊数、茎比密度和茎水分含量均呈减小趋势,并且种子重和种子短轴长也均呈减小的趋势;(3)对沿干旱梯度分布的4个酸枣种群而言,叶性状的平均变异系数(33.70%)枝性状的平均变异系数(32.41%)种子性状的平均变异系数(9.07%),并且酸枣性状间存在很强的协变。结果表明酸枣的地上部分形态性状沿干旱梯度表现出很强的变异,推测在未来的气候变化下,酸枣将通过这种表型变异的有效组合来适应环境变化。     

Artichoke Leaf Morphology and Surface Features in Different Micropropagation Stages

Artichoke (Cynara scolymus L.) leaf size and shape, glandular and covering trichomes, stomatal density, stomata shape, pore area and epicuticular waxes during micropropagation stages were studied by scanning electron microscopy (SEM) and morphometric analysis with the aim to improve the survival rate after transfer to greenhouse conditions. Leaves from in vitro shoots at the proliferation stage showed a spatular shape, ring-shaped stomata, a large number of glandular trichomes and juvenile covering hairs, but failed to show any epicuticular waxes. Leaves from in vitro plants at the root elongation stage showed a lanceolated elliptic shape with a serrated border, elliptical stomata, decreased pore area percentage, stomatal density, and mature covering trichomes. One week after transfer to ex vitro conditions, epicuticular waxes appeared on the leaf surface and stomata and pore area were smaller as compared to in vitro plants. Artichoke acclimatization may be improved by hormonal stimulation of root development, since useful morphological changes on leaves occurred during root elongation.     

Aegilops cylindrica var.kastorianum (Poaceae): A new variety from Greece

A native population ofAegilops cylindrica was encountered for the first time in Greece in 1980 (near Kastoria, NW. Greece), completely isolated and at a great distance from its main distribution area. There are morphological and chromosomal, but no protein and esterase pattern differences from otherAe. cylindrica populations. This justifies the recognition of a new variety: var.kastorianum. Our comparative and karyotypic observations support the view thatAe. caudata var.polyathera and not var.caudata (typica) is the possible donor of genome C of the new variety.     

Microsatellite analysis of the natterjack toad (Bufo calamita) in Denmark: populations are islands in a fragmented landscape

The European natterjack toad (Bufo calamita) has declined rapidly in recent years, primarily due to loss of habitat, and in Denmark it is estimated that 50% of the isolated populations are lost each decade. To efficiently manage and conserve this species and its genetic diversity, knowledge of the genetic structure is crucial. Based on nine polymorphic microsatellite loci, the genetic diversity, genetic structure and gene flow were investigated at 12 sites representing 5–10% of the natterjack toad localities presently known in Denmark. The expected heterozygosity (H _E) within each locality was generally low (range: 0.18–0.43). Further analyses failed to significantly correlate genetic diversity with population size, degree of isolation and increasing northern latitude, indicating a more complex combination of factors in determining the present genetic profile. Genetic differentiation was high (overall θ = 0.29) and analyses based on a Bayesian clustering method revealed that the dataset constituted 11 genetic clusters, defining nearly all sampling sites as distinct populations. Contemporary gene flow among populations was undetectable in nearly all cases, and the failure to detect a pattern of isolation by distance within major regions supported this apparent lack of a gene flow continuum. Indications of a genetic bottleneck were found in three populations. The analyses suggest that the remaining Bufo calamita populations in Denmark are genetically isolated, and represent independent units in a highly fragmented gene pool. Future conservation management of this species is discussed in light of these results. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Leaf morphology of a facultative rheophyte,Farfugium japonicum var.luchuense (Compositae)

The gross morphology and anatomy of leaves in a facultative rheophyte,Farfugium japonicum var.luchuense, were examined in order to clarify how the two characters are correlated with one another and how the facultative rheophyte differs from obligate rheophytes in leaf morphology and anatomy. Most rheophytes of the variety have narrowly cuneate leaf base, while most inland plants usually have truncate to cordate one, although the habitat-morphology correlation is not so clear. The leaf shape or the divergence angle of leaf base is strongly correlated with the number of primary veins and intervein-distance dependent on the number of cells intervening between veins. This makes a marked contrast to many reported cases of obligate rheophytes in which the leaf shape is strongly correlated with cell size. There is a rough tendency that narrower leaves of rheophytes have a thicker cuticular layer. However, the cell size and the volume (area) of intercellular space differ only slightly with the leaf shape.     

极小种群野生植物对开蕨种群结构特征和群落物种多样性

以国家Ⅱ级保护极小种群野生植物——对开蕨(Phyllitis scolopendrium)为研究对象,分析了在海拔729、1008m群落内,其种群大小、分布频度和密度,个体形态特征指标及其在种群内、种群间差异,苗高分布规律、相对苗高组替代年龄级结构,分布格局和群落各层次的物种多样性,群落的相似性。结果表明:对开蕨在自然分布区内为偶见种,呈斑块状分布。在400m~2内,海拔1008m处(01群落)种群密度为31株,样地分布频度为43.75%,最大密度15株/25m~2;海拔729m处(02群落)种群密度为91株,样地分布频度为93.75%,最大密度30株/25m~2。通过对其自然苗高,叶片数量,有孢子囊叶片数量,最大叶片长、宽值,最小叶片长、宽值,冠径,叶片厚度7个形态指标的分析显示,种群内变异较大,随着植株高度(年龄)的增加,其变异系数均随之减小而趋于稳定;种群间在自然苗高,最大叶片长度和宽度,成熟孢子叶片数量,冠径,叶片厚度上达到极显著差异(P0.01)。其苗高分布呈现双峰型,01种群波谷出现在15.1—20.0cm处;02种群波谷出现在10.1—15.0cm处,两群落均显示有一次较大的更新过程,同时01种群在40.1—45.0cm出现间断。年龄级分析得出,01种群划分为5个龄级,近于正态分布;02种群划分为4个龄级,为倒J型分布;两种群分别处于中龄期和幼龄期,没有出现衰退型年龄结构。格局分析得出,对开蕨种群均为聚集分布。两群落的乔木、灌木、草本层的Shannon-Wiener指数、Pielou均匀度指数、Marglef丰富度指数、生态优势度和种间相遇机率较低(与地带顶级植被相比)而且分布不均;对开蕨在两群落的草本层中重要值较低,仅为伴生种。两群落相似性分析显示,乔木、灌木、草本层的相似度指数分别为66.67%、69.23%和38.46%,草本层差异较大。     

Low levels of quantitative and molecular genetic differentiation among natural populations of Medicago ciliaris Kroch. (Fabaceae) of different Tunisian eco-geographical origin

In this paper, we analyze the genetic variability in four Tunisian natural populations of Medicago ciliaris using 19 quantitative traits and six polymorphic microsatellite loci. We investigated the amplification transferability of 30 microsatellites developed in the model legume M. truncatula to M. ciliaris. Results revealed that about 56.66% of analyzed markers are valuable genetic markers for M. ciliaris. The most genetic diversity at quantitative traits and microsatellite loci was found to occur within populations (>80%). Low differentiations among populations at quantitative traits Q _ST  = 0.146 and molecular markers F _ST  = 0.18 were found. The majority of measured traits exhibited no significant difference in the level of Q _ST and F _ST . Furthermore, significant correlations established between these traits and eco-geographical factors suggested that natural selection should be invoked to explain the level of phenotypic divergence among populations rather than drift. There was no significant correlation between population differentiation at quantitative traits and molecular markers. Significant spatial genetic structure consistent with models of isolation by distance was detected within all studied populations. The site-of-origin environmental factors explain about 9.07% of total phenotypic genetic variation among populations. The eco-geographical factors that influence more the variation of measured traits among populations are the soil texture and altitude. Nevertheless, there were no consistent pattern of associations between gene diversity (He) and environmental factors.     

Plagiochila cucullifolia var. anomala var. nov. from Ecuador, with notes on discordant molecular and morphological variation in Plagiochila

Plagiochila cucullifolia Jack & Steph. var. anomala J. Heinrichs & Gradst. var. nov. is described and illustrated. The new variety is known from a single locality in southern Ecuador and differs from P. cucullifolia var. cucullifolia by the flat, not saccate leaves, somewhat smaller plant size and weaker leaf dentation. According to phylogenetic analyses of 35 nrDNA ITS1 and ITS2 sequences of Plagiochila, the two varieties of P. cucullifolia form a monophyletic lineage and are placed in a well supported clade together with five other species of Plagiochila sect. Hylacoetes: P. dimorpha Lindenb. & Gottsche var. ecuadorica (Inoue) J. Heinrichs, P. flabelliflora Steph., P. patriciae J. Heinrichs & H. Anton, P. macrostachya Lindenb. and P. turgida Herzog. Within Plagiochila, nrITS sequence variation is not concordant with morphological diversification. ITS sequences of Plagiochila cucullifolia s.str. and of P. dimorpha var. ecuadorica differ in only 23 aligned positions whereas two sequences of P. subplana Lindenb. differ in 86 aligned positions. Morphologically, P. cucullifolia s.str. and P. dimorpha var. ecuadorica differ in more than 20 characters and previously these two taxa were placed in separate genera. P. subplana phenotypes show considerable variation in leaf shape and dentation but the extremes are linked by numerous intermediates.